Dynamic interplay of flow and collagen stabilizes primary hepatocytes culture in a microfluidic platform

Manjunath Hegde, Rohit Jindal, Abhinav Bhushan, Shyam Sundhar Bale, William J. McCarty, Inna Golberg, O. Berk Usta, Martin L. Yarmush

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

The creation of stable flow cultures of hepatocytes is highly desirable for the development of platforms for drug toxicity screening, bio-artificial liver support devices, and models for investigating liver physiology and pathophysiology. Given that hepatocytes cultured using the collagen overlay or in 'sandwich' configuration maintain a wide range of differentiated functions, we describe a simple method for adapting this culture configuration within a microfluidic device. The device design consists of a porous membrane sandwiched between two layers of PDMS resulting in a two-chambered device. In the bottom chamber, hepatocytes are cultured in the collagen sandwich configuration, while the top chamber is accessible for flow. We demonstrate that hepatocytes cultured under flow exhibit higher albumin and urea secretions and induce cytochrome P450 1A1 activity in comparison to static cultures. Furthermore, over two weeks, hepatocytes cultured under flow show a well-connected cellular network with bile canaliculi formation, whereas static cultures show formation of gaps in the cellular network that progressively increase over time. Although enhanced functional response of hepatocytes cultured under flow has been observed in multiple prior studies, the exact mechanism for this flow induced effect remains unknown. In our work, we identified that hepatocytes secrete a higher level of collagen in the flow cultures; inhibiting collagen secretion within the flow cultures reduced albumin secretion and restored the appearance of gaps in the cellular network similar to the static cultures. These results demonstrate the importance of the increased collagen secretion by hepatocytes cultured under flow as a mechanism to maintain a well-connected cellular network and a differentiated function.

Original languageEnglish (US)
Pages (from-to)2033-2039
Number of pages7
JournalLab on a Chip - Miniaturisation for Chemistry and Biology
Volume14
Issue number12
DOIs
StatePublished - Jun 21 2014
Externally publishedYes

Fingerprint

Microfluidics
Collagen
Hepatocytes
Lab-On-A-Chip Devices
Liver
Albumins
Bile Canaliculi
Artificial Liver
Equipment Design
Equipment and Supplies
Preclinical Drug Evaluations
Physiology
Drug-Related Side Effects and Adverse Reactions
Urea
Cytochrome P-450 Enzyme System
Toxicity
Screening
Membranes
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)
  • Bioengineering
  • Biomedical Engineering

Cite this

Dynamic interplay of flow and collagen stabilizes primary hepatocytes culture in a microfluidic platform. / Hegde, Manjunath; Jindal, Rohit; Bhushan, Abhinav; Bale, Shyam Sundhar; McCarty, William J.; Golberg, Inna; Usta, O. Berk; Yarmush, Martin L.

In: Lab on a Chip - Miniaturisation for Chemistry and Biology, Vol. 14, No. 12, 21.06.2014, p. 2033-2039.

Research output: Contribution to journalArticle

Hegde, M, Jindal, R, Bhushan, A, Bale, SS, McCarty, WJ, Golberg, I, Usta, OB & Yarmush, ML 2014, 'Dynamic interplay of flow and collagen stabilizes primary hepatocytes culture in a microfluidic platform', Lab on a Chip - Miniaturisation for Chemistry and Biology, vol. 14, no. 12, pp. 2033-2039. https://doi.org/10.1039/c4lc00071d
Hegde, Manjunath ; Jindal, Rohit ; Bhushan, Abhinav ; Bale, Shyam Sundhar ; McCarty, William J. ; Golberg, Inna ; Usta, O. Berk ; Yarmush, Martin L. / Dynamic interplay of flow and collagen stabilizes primary hepatocytes culture in a microfluidic platform. In: Lab on a Chip - Miniaturisation for Chemistry and Biology. 2014 ; Vol. 14, No. 12. pp. 2033-2039.
@article{25b3fdbc9ba4440aa33a45623605121b,
title = "Dynamic interplay of flow and collagen stabilizes primary hepatocytes culture in a microfluidic platform",
abstract = "The creation of stable flow cultures of hepatocytes is highly desirable for the development of platforms for drug toxicity screening, bio-artificial liver support devices, and models for investigating liver physiology and pathophysiology. Given that hepatocytes cultured using the collagen overlay or in 'sandwich' configuration maintain a wide range of differentiated functions, we describe a simple method for adapting this culture configuration within a microfluidic device. The device design consists of a porous membrane sandwiched between two layers of PDMS resulting in a two-chambered device. In the bottom chamber, hepatocytes are cultured in the collagen sandwich configuration, while the top chamber is accessible for flow. We demonstrate that hepatocytes cultured under flow exhibit higher albumin and urea secretions and induce cytochrome P450 1A1 activity in comparison to static cultures. Furthermore, over two weeks, hepatocytes cultured under flow show a well-connected cellular network with bile canaliculi formation, whereas static cultures show formation of gaps in the cellular network that progressively increase over time. Although enhanced functional response of hepatocytes cultured under flow has been observed in multiple prior studies, the exact mechanism for this flow induced effect remains unknown. In our work, we identified that hepatocytes secrete a higher level of collagen in the flow cultures; inhibiting collagen secretion within the flow cultures reduced albumin secretion and restored the appearance of gaps in the cellular network similar to the static cultures. These results demonstrate the importance of the increased collagen secretion by hepatocytes cultured under flow as a mechanism to maintain a well-connected cellular network and a differentiated function.",
author = "Manjunath Hegde and Rohit Jindal and Abhinav Bhushan and Bale, {Shyam Sundhar} and McCarty, {William J.} and Inna Golberg and Usta, {O. Berk} and Yarmush, {Martin L.}",
year = "2014",
month = "6",
day = "21",
doi = "10.1039/c4lc00071d",
language = "English (US)",
volume = "14",
pages = "2033--2039",
journal = "Lab on a Chip - Miniaturisation for Chemistry and Biology",
issn = "1473-0197",
publisher = "Royal Society of Chemistry",
number = "12",

}

TY - JOUR

T1 - Dynamic interplay of flow and collagen stabilizes primary hepatocytes culture in a microfluidic platform

AU - Hegde, Manjunath

AU - Jindal, Rohit

AU - Bhushan, Abhinav

AU - Bale, Shyam Sundhar

AU - McCarty, William J.

AU - Golberg, Inna

AU - Usta, O. Berk

AU - Yarmush, Martin L.

PY - 2014/6/21

Y1 - 2014/6/21

N2 - The creation of stable flow cultures of hepatocytes is highly desirable for the development of platforms for drug toxicity screening, bio-artificial liver support devices, and models for investigating liver physiology and pathophysiology. Given that hepatocytes cultured using the collagen overlay or in 'sandwich' configuration maintain a wide range of differentiated functions, we describe a simple method for adapting this culture configuration within a microfluidic device. The device design consists of a porous membrane sandwiched between two layers of PDMS resulting in a two-chambered device. In the bottom chamber, hepatocytes are cultured in the collagen sandwich configuration, while the top chamber is accessible for flow. We demonstrate that hepatocytes cultured under flow exhibit higher albumin and urea secretions and induce cytochrome P450 1A1 activity in comparison to static cultures. Furthermore, over two weeks, hepatocytes cultured under flow show a well-connected cellular network with bile canaliculi formation, whereas static cultures show formation of gaps in the cellular network that progressively increase over time. Although enhanced functional response of hepatocytes cultured under flow has been observed in multiple prior studies, the exact mechanism for this flow induced effect remains unknown. In our work, we identified that hepatocytes secrete a higher level of collagen in the flow cultures; inhibiting collagen secretion within the flow cultures reduced albumin secretion and restored the appearance of gaps in the cellular network similar to the static cultures. These results demonstrate the importance of the increased collagen secretion by hepatocytes cultured under flow as a mechanism to maintain a well-connected cellular network and a differentiated function.

AB - The creation of stable flow cultures of hepatocytes is highly desirable for the development of platforms for drug toxicity screening, bio-artificial liver support devices, and models for investigating liver physiology and pathophysiology. Given that hepatocytes cultured using the collagen overlay or in 'sandwich' configuration maintain a wide range of differentiated functions, we describe a simple method for adapting this culture configuration within a microfluidic device. The device design consists of a porous membrane sandwiched between two layers of PDMS resulting in a two-chambered device. In the bottom chamber, hepatocytes are cultured in the collagen sandwich configuration, while the top chamber is accessible for flow. We demonstrate that hepatocytes cultured under flow exhibit higher albumin and urea secretions and induce cytochrome P450 1A1 activity in comparison to static cultures. Furthermore, over two weeks, hepatocytes cultured under flow show a well-connected cellular network with bile canaliculi formation, whereas static cultures show formation of gaps in the cellular network that progressively increase over time. Although enhanced functional response of hepatocytes cultured under flow has been observed in multiple prior studies, the exact mechanism for this flow induced effect remains unknown. In our work, we identified that hepatocytes secrete a higher level of collagen in the flow cultures; inhibiting collagen secretion within the flow cultures reduced albumin secretion and restored the appearance of gaps in the cellular network similar to the static cultures. These results demonstrate the importance of the increased collagen secretion by hepatocytes cultured under flow as a mechanism to maintain a well-connected cellular network and a differentiated function.

UR - http://www.scopus.com/inward/record.url?scp=84901051242&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84901051242&partnerID=8YFLogxK

U2 - 10.1039/c4lc00071d

DO - 10.1039/c4lc00071d

M3 - Article

C2 - 24770663

AN - SCOPUS:84901051242

VL - 14

SP - 2033

EP - 2039

JO - Lab on a Chip - Miniaturisation for Chemistry and Biology

JF - Lab on a Chip - Miniaturisation for Chemistry and Biology

SN - 1473-0197

IS - 12

ER -